Accurate apoptosis measurement requires quantification of loss of expression of surface antigens and cell fragmentation

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
David DiazMelchor Alvarez-Mon

Abstract

The use of ratiometric cell enumeration methods emerges as a more accurate method of measurement of the occurrence of apoptosis in cell cultures. These new flow cytometry methods were used to quantify the impact of cell fragmentation and loss of lineage antigen (LAg) expression on measurement of apoptosis. Highly purified human lymphocyte populations were negatively sorted and cultured for 24 h. Apoptotic cells were identified using annexin V, 7-amino-actinomycin D and their LAgs were stained with antibodies. A new indicator, the apoptotic rate, was used to determine apoptosis occurrence and its validity compared with the widely accepted percentage of apoptotic cells (apoptotic index, AI). Loss of LAg expression and cell fragmentation were observed under all conditions assayed and for all cell populations studied. Current methods for quantifying of apoptosis involving AI systematically underestimate apoptosis occurrence in all populations and conditions, especially among cells undergoing spontaneous apoptosis.

References

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Apr 15, 1999·Cellular Immunology·A PotterP S Rabinovitch
Oct 31, 2002·Nature Immunology·Hung T Khong, Nicholas P Restifo
Jun 5, 2004·Journal of Leukocyte Biology·D DiazM Alvarez-Mon

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Citations

Mar 14, 2014·British Journal of Haematology·Ulrika JohanssonUNKNOWN British Committee for Standards in Haematology
Apr 27, 2007·PloS One·Daniel StockholmAndras Paldi
Jan 4, 2007·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Andrea Lisa HolmeShazib Pervaiz

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Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis